13C NMR Spectroscopy

Introduction
• Nuclear magnetic resonance spectroscopy (NMR) is the most
powerful tool available for structural determination.
• A nucleus with an odd number of protons, an odd number of
neutrons, or both, has a nuclear spin that can be observed by the
NMR spectrometer.
• NMR active nuclei include: 1H, 13C, 19F, and 31P.
• Remember a spinning nucleus generates a magnetic field (magnetic
moment).
• In the absence of an external magnetic field, proton magnetic
moments have random orientations.
• However, in the presence of an external magnetic field, the
magnetic moment is aligned either with or against the external field.

2
 Introduction

• The stronger the magnetic field, the greater the energy difference
between the two spin states, resulting in a greater population
difference between the two states greater sensitivity.

3
 Nuclear Spin Energy Levels
• A photon of light with the right amount of energy (radiofrequency, rf)
can be absorbed and cause the spinning proton to flip.

External magnetic field

E2 E2

h
absorption
of energy

E1 E1

• The nuclei undergo a “spin flip”, and the nuclei are said to be “in
resonance”. This absorption of energy leads to the NMR signal

4
 Nuclear Spin Energy Levels

• If the two states become equally populated, then no net

spin transitions occur and no signal is produced. This is
called saturation.
• The frequency of EM radiation necessary for resonance
depends on the strength of the magnetic field and on the
chemical environment of the nucleus.
• Fortunately, protons (in 1H NMR) in molecules usually
experience different chemical environments (i.e. are
shielded to varying extents).

5
 1H NMR Spectroscopy

• Therefore, different frequencies are required to bring

different protons into resonance.
• Consider CH3OH:
H
Deshielded, senses higher Shielded, senses a smaller
effective magnetic field so O C H effective magnetic field so
comes into resonance at a H comes into resonance at a
higher frequency. H lower frequency.

6
 1H NMR Spectroscopy
• A 1H NMR spectrum provides the following information:
1. The # of different types of H – number of basic groups of
signals.
2. The relative numbers of different types of H –
3. The electronic environment of the different types of H –

4. The number of hydrogen “neighbors” a proton has –

7
 Simple Correlation Table of 1H
Chemical Shifts

* See text and Lab manual for more extensive tables 8

 Why Carbon (13C) NMR Spectroscopy
O

• Some organic compounds have few C-H bonds: HO

C
C
C O
C
C
HO
O
• Others have very similar 1H NMR spectra:

O O
H3C CH3 H3C H

H H H CH3
O O

2,6-dimethylbenzoquinone 2,5-dimethylbenzoquinone 9
 Carbon (13C) NMR vs 1H NMR
• The 13C nucleus can also undergo nuclear magnetic resonance.
• 13C NMR vs 1H NMR :

– 12C, the most abundant isotope of carbon, does NOT exhibit

NMR behavior. Why?
– 13C, only _____ natural abundance, does exhibit NMR behavior.

– Due to low abundance, 13C-13C coupling is usually not observed.

– Chemical shift ranges are much larger –
– Integration in 13C NMR is NOT reliable due to variable relaxation
times from C to C. Also Nuclear Overhauser Effect - the
intensity of the C signal increases as the number of attached
protons increases. Not uniform however.

10
 Fourier Transform (FT) spectroscopy
• The magnetic moment of the 13C nucleus is about 1/4 that of the H
nucleus resulting in lower sensitivity.
• The low natural abudance and small magnetic moment of the 13C
isotope results in the 13C nucleus being about ______ less sensitive
than the 1H nucleus to NMR phenomena.
• Consequently, much longer acquisition times were required.
• The development of Fourier transform (FT) spectroscopy has
made 13C NMR acquisition routine.
• The old way of acquiring NMR was to apply a constant magnetic
field to the sample and scan the range of frequencies = continuous
wave (CW) NMR.
• With FT-NMR the data is collected all at once by exciting the sample
with an RF pulse (typically only a few microseconds long) which
covers all the resonance frequencies, and thus changes the
orientation of all the protons.
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 Fourier Transform (FT) spectroscopy
• After the pulse has stopped, the decay of the signal from the sample
is measured. The decaying sine wave called a free induction
decay (FID):

O
Intensity of signal

H3C C CH3
Fourier
Transform

Time (s) Frequency

• A Fourier transform converts the intensity vs time data into

intensity vs frequency information.

12
Fourier Transform (FT) spectroscopy

Fourier
Transform

13
 Chemical Shifts in 13C NMR

• Two simple ideas will make interpretation of 13C

NMR spectra easier:
1. Hybridization of the C atom determines the
chemical shift:
 sp3 hybridized carbons have chemical shift
values _________.
 sp2 hybridized carbons have chemical shift
values _________.
2. The presence of an EN element near a C atom will
cause its chemical shift to move _____________.

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 Simple Correlation Table of 13C
chemical shifts

See text (p. 593) and Lab manual (p. 60) for more extensive tables

15
 Coupling in Carbon NMR
• The low abundance of 13C makes C-C coupling very rare.
• However, 13C-H coupling is common. N+1 rule still applies:

Coupling constants are large ~100-200 Hz for directly attached H’s. 16

 Coupling in Carbon NMR
• Spectra which show 13C-H coupling are called proton-
coupled spectra.
• However, extensive 13C-H coupling often produces
splitting patterns that are difficult to interpret.
• To simply 13C NMR spectra, often recorded using broad
band proton decoupling.
• Therefore each carbon signal appears as a singlet,
because C-H splitting has been eliminated.
• Spectra recorded in the broad band proton decoupling
mode give the number of unique carbon atoms in a
molecule.

17
Proton-coupled vs Proton-decoupled
13C NMR Spectra

18
 Interpreting 13C NMR Spectra
CH3
H3C C CH2 CH3
CH3

CDCl3
solvent
TMS

19
Interpreting 13C NMR Spectra

O
CH3 C O CH2 CH3

20
Interpreting 13C NMR Spectra

CH3

21
Interpreting 13C NMR Spectra
O O
H3C CH3 H3C H

H H H CH3
O O

22
Mass Spectrometry
 Basic Principles
• Mass Spectroscopy (MS) is a destructive analytical technique for
measuring the ______________ (____) of ions in the gas phase.

 This allows accurate determination of the _______________ of a

molecule.
 Structural information is also gained.
 Molecular Formula determination is sometimes possible.

• While the method is destructive, only very small amounts (1 mg or

less) is required.

24
 Basic Principles
• MS does not involve the absorption or emission of light.
• A mass spectrometer is designed to do 3 things:
1. Convert a neutral molecule, M, into positive (or negative) ions
usually by bombardment with a beam of high energy
electrons.

M + e M + 2e
10-70 eV
in energy
1 eV = 23 kcal/mol

2. Separate the ions based on mass (mass-to-charge ratio, ___).

3. Measure the relative abundance of each ion.

25
 Schematic of Mass Spectrometer
• First the sample is vaporized under vacuum.
• A beam of electrons bombardsElectron Impact in the gas phase
the molecules
Ionization
causing ionization and formation Source
of radical cations.
~70 Volts

Electron Collector (Trap)

Electron impact
Ionization source Positive
Ions
+
Repeller Neutral Inlet _
_
Molecules
+ + + to
+ + + +
Analyzer

e- e- e-
_
Electrons

Filament Extraction
Plate

26
 Schematic of Mass Spectrometer
• The radical cations fragment further after ionization owing to the
large amount of energy transferred by the electron beam.
• Some fragments carry a positive charge, others are neutral:

[CH3] + H
H m/z =
-e
CH4 H C H
H
m/z = [CH2] + 2H
m/z =
Molecular ion

• Only the positively charged fragments are accelerated into the

analyzer tube.

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 Schematic ofSector
Magnetic Mass MassSpectrometer
Analyzer

• The analyzer tube is surrounded by a magnet whose magnetic field

deflects the positively charge fragments in a curved path.
ion trajectory
in register
ion trajectory
not in register
(too light)

Ion Detector
Source
N ion trajectory
not in register
Electromagnet (too heavy)

• The amount of deflection depends on m/z.

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 Basis of Fragment Separation
• Fragments with smaller m/z value are deflected ______ than a
larger m/z value.
• Since z is usually _____, the fragments are sorted by mass.
• By varying the magnetic field, cations of different masses are sorted
and counted by a detector.
• The more stable the fragment the more likely it will make it to the
detector.
• The masses are graphed or tabulated according to their relative
abundance = The Mass Spectrum.

29
The Mass Spectrum of Methane
Base peak
m/z Intensity
1 3.4
2 0.2
M+ = 15 M+ = 16
12 2.8 C12H3+ Molecular ion
[C12H4]+.
13 8.0
[C12H2]+.
14 16.0
15 86.0 C12H+
16 100.0 [C12]+.
17 1.11
12 13 14 15 16 17
m/z 30
 Isotopes
• Most elements common to organic compounds are mixtures of isotopes.
• The existence of atomic isotopes in nature accounts for the appearance of
M+1 and M+2 peaks in a mass spectrum.
• Organic compounds containing only C, H, O, and N usually have relatively
small M+1 and M+2 peaks.

M+ M+
Relative abundance, %

M+1+
M+1+ M+2+

m/z m/z 31
C6H12 C20H42
 Isotopes
Element Most abundant isotope Less abundant isotope Relative abundance

Hydrogen 1H 2H 0.016

Carbon 12C 13C 1.08

Nitrogen 14N 15N 0.38

Oxygen 16O 18O 0.20

Sulfur 32S 34S 4.4

Chlorine 35Cl 37Cl 32.5

Bromine 79Br 81Br 98.0

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 Isotopes
• MS is particularly valuable for compounds which contain Cl and Br:
 If one S atom is present, M + 2 is ~ 4% of M+.
 If one Cl atom is present, M + 2 is ~ 33% of M+.
 If one Br atom is present, M + 2 is ~ to M+.
M 24 M 3 M 1
M+2 1 M+2 1 M+2 1

M+
Relative abundance, %

M+ M+ M+2+

M+2+

M+2+
33
m/z m/z m/z
Mass Spectrum with Chlorine

Cl
H3C CH CH3

M+2+

M+

34
Mass Spectrum with Bromine

H3C CH2 CH2 Br

M+ M+2+

35
 Isotopes
• Carbon Rule – For compounds containing only C, H, and O, the
following formula can be used to determine the number of carbons
in the molecule:
relative intensity of M + 1 peak
no. C s =
1.1

Determine the molecular formula of the unknown organic compound

whose mass spectral data is given in the table below:
Peak Mass Relative
(m/z) intensity
M 86 100.0
M+1 87 5.6
M+2 88 0.4

36
 Isotopes
• Nitrogen Rule: if a compound has:
– An odd number of nitrogen atoms, its molecular ion, M+, will be
odd.
– Zero or an even number of nitrogen atoms, its molecular ion, M+,
will be even.

37
 Resolution
• Resolution: a measure of how well a mass spectrometer separates
ions of different mass.
 Low resolution – capable of distinguishing among ions of different
nominal mass, i.e. different by at least one or more amu.
 High resolution – capable of distinguishing among ions that differ in
mass by as little as 0.0001 amu.

• For example: CO, N2, and ethene all have a nominal mass of 28
amu. High resolution MS can distinguish these molecules.

CO 27.9949 amu
N2 28.0061 amu
CH2=CH2 28.0314 amu

38
 Fragmentation Pathways
• Structural information is available from analysis of fragments formed
by bond cleavages in the molecular ion, M+.
• In general, the molecular ion, M+, will fragment so as to form the
most stable cationic fragment (usually a carbocation).
• In some cases, the M+ peak is very small or absent. Occurs if the
fragments are considerably more stable M+.

39
 Mass Spectrum-Fragmentation
• Consider the mass spectrum of pentane: p. 516-517 text

• Fragmentation of the molecular ion often results:

40
Mass Spectrum of Pentane

41
Fragmentation of Alkanes
43 CH3
CH3 CH2 CH2 CH CH3

71

57
86 M+

CH3
CH3 CH2 CH2 + HC CH3
1
m/z 43
CH3 CH3
2
CH3 CH2 CH2 CH CH3 CH3 CH2 CH2 CH + CH3

3 m/z 71
m/z 86 CH3
CH3 CH2 + H2C CH CH3
m/z 57
42
 Compounds with Heteroatoms
• Molecules containing O, N, halogens, or other heteroatoms often
undergo ___________ (adjacent to heteroatom).
• Driving force is resonance stabilized cations.

43
 Fragmentation of Alcohols
• Alcohols common fragmentation is -cleavage and loss of “H2O” to
give an M-18 peak.

M-29
OH

H3C C CH2 CH3

CH3

M+ = 88 (not observed) M-15

M-18 -15
M-18

44
Fragmentation of Amines
+
CH2=NH2
m/z=30 H3C
CH CH2 NH2
H3C

M+ = 73

Mass spectrum of isobutylamine

45
Fragmentation of Ketones

43

H3C C CH2 CH3

M+ = 72

57

Mass spectrum of 2-butanone

46
 McLafferty Rearrangement
• If one of the alkyl groups attached to the carbonyl carbon of an
aldehyde or ketone has a hydrogen, a cleavage known as a
McLafferty rearrangement can occur.
Mass spectrum of butyraldehyde

M–28
O

M+=72

47
 Aromatic Compounds
• Usually strong M+ peak.
• m/z =91 for tropylium ion and methylene spacings above 91 (105,
119, etc. for alkyl chains) often observed.
• m/z = 65 (C5H5+), 77 (C6H5+) are sometimes observed.

CH2 R CH2
-R

m/z 91

m/z 91

48
Aromatic Compounds

49
 Common Fragments
O O
CH3 CH 3CH2 C C
H H3C
m/z = 15 29 29 43

O
O CH2 C
C
HO

m/z = 45 91 105

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